ALG10 Alleviates IBD-Related Intestinal Fibrosis by Modulating the TGF-β1/PI3K/Akt Signaling Pathway

To explore the role of asparagine-linked glycosylation 10 (ALG10) in intestinal fibrosis (IF) and elucidate its underlying molecular mechanisms.

Western blot and RT-qPCR were employed to detect ALG10 expression in a TNBS/DSS-induced mouse IF model and in primary colonic fibroblasts. TNBS/DSS-induced ALG10^-/- mouse models were established, and primary colonic fibroblasts were isolated in vitro. Pathological changes and collagen deposition in colon tissues were evaluated via Masson's trichrome staining and H&E staining. RT-qPCR, Western blot, and ELISA were performed to measure the expression of IF-related markers. A TGF-β1-induced CCD-18Co cell model was established, and immunofluorescence was used to detect the fluorescence intensity of COL I and E-cadherin. The interaction mechanism between ALG10 and TGF-β1 was further explored by molecular docking, MD simulation, Co-IP and dual-luciferase reporter gene experiments.

Compared with the WT group, ALG10 expression was significantly increased in the TNBS/DSS group. In both in vivo and in vitro models, ALG10 silencing markedly alleviated fibrosis severity and pathological damage, downregulated the expression of COL I and α-SMA, and upregulated E‑cadherin expression and significantly reduced the levels of p‑PI3K, p‑Akt, and the target genes FGF2 and p‑FoxO1/3a. Molecular docking, MD simulations, and Co-IP assays suggested that ALG10 potentially interacts with TGF-β1, while dual-luciferase reporter assays further demonstrated that ALG10 promotes TGF-β1 promoter activity.

ALG10 acts as a key mediator promoting the progression of IF. Silencing ALG10 can alleviate IBD-related IF, a mechanism that may involve the regulation of the TGF‑β1/PI3K/Akt signaling pathway.